PROJECT SUMMARY/ABSTRACT Project Title: Scientific Core 2: Protein Expression and Characterization Core We will establish and operate a protein expression and characterization core that serves the Cooperative Research Center for Nano-Scaffold Vaccines Against Chlamydia trachomatis (Ct) by providing facilities, expertise, effort, materials, and information management to help achieve the aims of the individual projects. We will establish and maintain facilities with lab space, instrumentation, and storage necessary for production, characterization, preservation, and distribution of expression plasmids, nanolipoprotein particles, and protein antigens. We will establish and maintain procedures and training materials sufficient to develop personnel with the expertise necessary to support the protein production needs of the individual projects. The facility will be staffed to provide the effort required for protein production and characterization, as requested by participating projects. We will operate as an on-demand service center, although commonly used, archivable materials may be produced in bulk, aliquoted, and stored for ready distributions. A regular production schedule may also be necessary for perishable materials that form the basis of a projects' vaccine technology, plasmids for cell-free translation within Project 1, and fully formed NLPs for RNA-based vaccines in project 2, for example. For regularly used materials, we will establish quality metrics and maintain a quality control program to ensure reproducibility of results from materials we generate. We will work with affiliated projects to aggregate production and characterization protocols and to determine acceptable tolerances for metrics of quality control. We will establish an information management system, likely rudimentary to start, such that materials distributed from the core can be traced back to the procedures, personnel, materials, and associated metadata, such as quality assessments, associated with the distributed materials across all 3 Projects. The core will be highly centralized with personnel and facilities at Lawrence Livermore National Laboratory (LLNL), but some key equipment and expertise may not be available at LLNL, in which case we will partner with outside facilities. As time and resources permit, the protein production and characterization core will work to diversify the portfolio of nanolipoprotein particles for vaccine delivery, scale production of NLPs, formulate less perishable preparations of NLPs, and explore use of single particle methods, such as particle tracking, for characterization and quality control. The protein production and characterization core can also work as a user facility, providing training and access to facilities that might enable research and development for individual project that do not have the expertise or equipment at their locations.